MSC-Derived Exosomes and MicroRNA in Glioma Therapy

MSC 衍生的外泌体和 MicroRNA 在神经胶质瘤治疗中的应用

• 批准号: 10305631
• 负责人: FREDERICK F LANG
• 金额: $ 35.87万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-11 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10305631
• 关键词: Address; Adult; Biology; Blood; Brain; Brain Neoplasms; Clinic; Data; Disease; Dose; Down-Regulation; Fibroblasts; Gene Expression; Genes; Glioblastoma; Glioma; Goals; Grant; Growth; Home; Homing; Human; In Vitro; Lentivirus; Light; Literature; Malignant neoplasm of brain; Mass Spectrum Analysis; Mediating; Mesenchymal Stem Cells; Methods; MicroRNAs; Mus; New Agents; Patients; Persons; Primary Brain Neoplasms; Proteins; Proteomics; Public Health; Regulator Genes; Reporter; Safety; Schedule; Surface; System; Testing; Tetracyclines; The Cancer Genome Atlas; Therapeutic; Therapeutic Uses; Time; Translations; Tumor Suppressor Proteins; Untranslated RNA; base; bioluminescence imaging; blood-brain tumor barrier; bone marrow mesenchymal stem cell; delivery vehicle; differential expression; effective therapy; efficacy testing; exosome; high throughput screening; improved; in vivo; in vivo bioluminescence imaging; nanovesicle; novel; novel strategies; novel therapeutic intervention; overexpression; prevent; restoration; stem cell exosomes; stem-like cell; therapeutic miRNA; therapeutically effective; therapy resistant; treatment response

Glioblastoma (GBM), the most common and deadly adult primary brain tumor, is notoriously resistant to therapy, not only because of its unique biology, including Glioma Stem-like Cells (GSCs), but also because of the delivery challenges imposed by the blood-brain and blood-tumor barriers. Thus, there is an urgent need to identify effective GBM-specific therapeutics and to elucidate strategies for delivering these new agents. A promising new approach is treatment with microRNAs (miRs), which are small, noncoding RNAs that are powerful regulators of gene expression. We have shown that restoration of tumor suppressor miRs is capable of killing GSCs and, therefore, treatment with miRs is potentially a new paradigm in GBM therapy. However, it is currently unknown which miRs will be most effective against GBMs and how these miRs should be delivered. This grant is significant because it directly addresses both of these problems by identifying the most effect anti-GBM miRs and by developing exosomes derived from bone marrow mesenchymal stem cells (MSCs) as a new delivery strategy for these miRs. Exosomes are naturally occurring nanovesicles that function as intercellular transport vehicles. We hypothesize that miRs can be used as effective therapeutics against GBMs, and that exosomes derived from cultured MSCs can be used to systemically deliver these antiglioma miRs. Aim 1 will identify miRs that are most effective against GSCs. In an in vitro high-throughput screen (HTS) of 578 miRs against a panel of GSCs, we have already identified 50 antiglioma miRs that effectively kill GSCs in vitro. This aim will use an in vivo HTS to determine which of the 50 top candidates is most effective against orthotopic GSCs in mice, and it will define the most effective miR combinations. Mechanistic studies will identify the target(s) responsible for the anti-GBM effects. Aim 2 will develop MSC-derived exosomes as novel delivery vehicles of antiglioma miRs. We have shown that after transduction with lentiviruses containing antiglioma miRs, MSCs package the miRs into exosomes (called Exos-miRs) and secrete the Exo-miRs into the culture medium. These Exo-miRs can be isolated and delivered systemically, after which they home to GSCs and inhibit target genes. This aim will define the optimal dose/schedule of Exos-miRs using bioluminescence imaging and a novel TET-reporter system. This dose/schedule will be used to test the efficacy and safety of the best antiglioma miRs or miR combinations identified in Aim 1. Aim 3 will elucidate the mechanisms underlying the ability of Exos-miRs to home to GBMs. By using proteomic analyses to compare strongly homing exosomes (MSC-Exos) with poorly homing ones (fibroblast-Exos), we have identified candidate homing proteins. In this aim, we will perform functional analyses to determine the extent to which the identified proteins mediate the homing of Exo-miRs to GBMs. Successful completion of this grant will result in a new and integrated agent/delivery paradigm that uses miRs to treat GBMs and MSC-derived exosomes to deliver these miRs, and that has the potential to improve the survival of the estimated 15,000 people who die of GBM each year due to the current lack of effective therapy for this disease.

胶质母细胞瘤（GBM）是最常见，最致命的成人原发性脑肿瘤，众所周知，对治疗具有抗药性 不仅是因为其独特的生物学，包括神经胶质瘤干细胞（GSC），还因为递送 血脑和血肿瘤障碍施加的挑战。因此，迫切需要确定 有效的GBM特异性治疗疗法，并阐明提供这些新代理的策略。一个有希望的新 方法是用microRNA（miRS）处理，它们是小的，非编码的RNA，是强大的调节剂 基因表达。我们已经表明，肿瘤抑制miR的恢复能够杀死GSC，并且， 因此，用miR治疗可能是GBM治疗中的新范式。但是，目前未知 哪些mir对GBM最有效，以及这些mir应该如何传递。这笔赠款很重要 因为它通过识别抗GBM mir和通过 开发源自骨髓间充质干细胞（MSC）作为新的递送策略的外泌体 对于这些mir。外泌体是自然发生的纳米孢子，它们充当细胞间运输车辆。 我们假设MIR可以用作针对GBM的有效治疗剂，并且外泌体来自 培养的MSC可用于系统地传递这些抗激瘤miR。 AIM 1将识别最多的mir 对GSC有效。在与GSC面板的578 mir的体外高通量屏幕（HTS）中，我们 已经鉴定出50种在体外有效杀死GSC的抗糖瘤miR。这个目标将使用体内HTS 确定50个顶级候选者中的哪个最有效地针对小鼠的原位GSC，它将定义 最有效的miR组合。机械研究将确定负责抗GBM的靶标 效果。 AIM 2将发展为MSC衍生的外泌体作为抗激瘤miR的新型递送车。我们有 表明，用含有抗糖尿瘤miR的慢病毒转导后，MSC将miR贴入 外泌体（称为Exos-Mirs），并将外来型分泌到培养基中。这些Exo-Mirs可以是 孤立和全身传递，之后他们回到GSC并抑制靶基因。这个目标将定义 使用生物发光成像和新型TET-Reporter系统的Exos-Mirs的最佳剂量/时间表。这 剂量/时间表将用于测试最佳抗激瘤miR或miR组合的功效和安全性 在AIM 1中确定的AIM 3将阐明Exos-Mirs在家中GBM的能力的基础机制。 通过使用蛋白质组学分析来比较强烈的归巢外泌体（MSC-exos）与归巢较差 （成纤维细胞-EXO），我们已经确定了候选归巢蛋白。在此目标中，我们将执行功能分析 确定鉴定蛋白质介导exo-mirs向GBM的归巢的程度。成功的 该赠款的完成将导致一个新的综合代理/交付范式，该范式使用mirs来治疗GBMS 和MSC衍生的外泌体提供这些miR，这有可能提高 由于目前缺乏对这种疾病的有效治疗，估计每年死于GBM的15,000人。